EFSA Journal 2015;13(5):4085 SCIENTIFIC OPINION Scientific Opinion on re-evaluation of chlorophyllins (E 140(ii)) as food additives1 EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS)2,3 European Food Safety Authority (EFSA), Parma, Italy ABSTRACT Chlorophyllins (E 140(i)) are obtained by saponification of a solvent extract from sources, such as grass, lucerne, and nettle, that could not be regarded as edible plant material or food for humans. Chlorophyllins represent 90 % of the colouring matter in the food additive E 140(ii); the remaining part consists of other pigments, such as carotenoids, together with oils, fats and waxes derived from the source material. The Panel noted that the material used in many studies, identified as “chlorophyllins”, was quite often, if not always, a copper complex of chlorophyllins (E 141(ii)). There are no data regarding the absorption, distribution, metabolism and excretion (ADME) and toxicity of chlorophyllins (E 140(ii)). Considering the available data on chlorophylls (E 140(i)), the Panel concluded that chlorophyllins are not metabolites of chlorophylls in humans and owing to their differences in physico-chemical properties, it was not possible to support read-across for toxicity data between these two compounds. The Panel considered that it is necessary to carefully review the definition and identity of E 140(ii) in order to adequately characterise the food additive E 140(ii)) as used in the market. This will also allow proper assessment of its safety when relevant studies of the compound to which consumers are actually exposed become available. Considering the absence of relevant ADME and toxicity data, and because chlorophyllins (E 140(ii)) are neither natural constituents of the regular diet nor metabolites of chlorophylls in humans, the Panel concluded that it was not possible to assess the safety of chlorophyllins (E 140(ii)) as food additives. © European Food Safety Authority, 2015 KEY WORDS Chlorophyllins, chlorophyllin a, chlorophyllin b, chlorin e6, rhodin g7, E 140(ii), food colours 1 On request from the European Commission, Question EFSA-Q-2011-00371 adopted on 15 April 2015. 2 Panel members: Fernando Aguilar, Riccardo Crebelli, Alessandro Di Domenico, Birgit Dusemund, Maria Jose Frutos, Pierre Galtier, David Gott, Ursula Gundert-Remy, Claude Lambré, Jean-Charles Leblanc, Oliver Lindtner, Peter Moldeus, Alicja Mortensen, Pasquale Mosesso, Agneta Oskarsson, Dominique Parent-Massin, Ivan Stankovic, Ine Waalkens- Berendsen, Rudolf Antonius Woutersen, Matthew Wright and Younes Maged. Correspondence: [email protected] 3 Acknowledgement: The Panel wishes to thank the members of the former Working Group “A” Food Additives and Nutrient Sources (2011-2014) and the members of the Standing Working Group on the re-evaluation of food colours: Fernando Aguilar, Riccardo Crebelli, Alessandro Di Domenico, Maria Jose Frutos, Pierre Galtier, David Gott, Claude Lambré, Jean-Charles Leblanc, Agneta Oskarsson, Jeanne Stadler, Paul Tobback, Ine Waalkens-Berendsen and Rudolf Antonius Woutersen, for the preparatory work on this scientific opinion and EFSA staff members: Federica Lodi, Ana Maria Rincon and Alexandra Tard for the support provided to this scientific opinion. EFSA acknowledges those European competent authorities, food industry and other stakeholders that provided occurrence data (usage and analytical data) on food additive (E 140(ii)) in food and beverages, and supported the data collection for the Comprehensive European Food Consumption Database. Suggested citation: EFSA ANS Panel (EFSA Panel on Food Additives and Nutrient Sources added to Food), 2015. Scientific Opinion on the re-evaluation of chlorophyllins (E 140(ii)) as food additives. EFSA Journal 2015;13(5):4085, 42 pp. doi:10.2903/j.efsa.2015.4085 Available online: www.efsa.europa.eu/efsajournal © European Food Safety Authority, 2015 Re-evaluation of chlorophyllins (E 140(ii)) as food additives SUMMARY Following a request from the European Commission (EC), the Panel on Food Additives and Nutrient Sources added to Food (ANS) was asked to deliver a scientific opinion re-evaluating the safety of chlorophyllins (E 140(ii)) when used as food additives. The Panel was not provided with a newly submitted dossier and based its evaluation on previous evaluations and additional literature that has become available since then. No new toxicological or biological information was submitted to the Panel for the re-evaluation of chlorophyllins (E 140(ii)) following European Food Safety Authority (EFSA) public calls for data. The Panel noted that not all of the original studies on which previous evaluations were based were available for this re-evaluation. To assist in identifying any emerging issue or any information relevant for the risk assessment, EFSA outsourced a contract to deliver an updated literature review on toxicological endpoints, dietary exposure and occurrence levels of chlorophyllins (E 140(ii)), which covered the period up to the end of 2014. Chlorophyllins (E 140(ii)) are authorised as food additives in the European Union (EU) in accordance with Annex II to Regulation (EC) No 1333/2008. The Panel noted that in this regulation, chlorophylls and chlorophyllins are authorised with the same E number, E 140. However, according to Commission Regulation (EU) No 231/2012, separate specifications are defined for chlorophylls (E 140(i)) and chlorophyllins (E 140(ii)). The Panel decided to re-evaluate these two food additives separately, given their different physico-chemical properties. Chlorophyllins (E 140(ii)) do not appear to have been evaluated previously by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) and the Scientific Committee on Food (SCF). Alkali salts of chlorophyllins are obtained by saponification of a solvent extract from sources, such as grass, lucerne and nettle, that could not be regarded as regular edible plant material or food for humans. Chlorophyllins represent 90 % of the colouring matter in the food additive E 140(ii); the remaining part consists of other pigments, such as carotenoids, together with oils, fats and waxes derived from the source material. According to the Natural Food Colours Association (NATCOL, 2011d): “The levels for residual solvent require two revisions to accommodate for the regular presence of ethanol and methanol at levels that are not caused by the use as solvents but result from the carryover from raw materials or reactions during extraction and processing”. The Panel also considered that the raw material should fulfil the conditions of current regulation (Commission Regulation (EU) No 231/2012) as regards maximum levels for possible contaminants, including pesticides residues applied during cultivation and mycotoxins. The Panel noted that, according to NATCOL (2011c) “magnesium chlorophyllin is not an article of commerce and the main constituents of the material of commerce are chlorin e6 and rhodin g7”. This is in contradiction with the main constituents of E 140(ii) as defined in Commission Regulation (EU) No 231/2012. The Panel noted that the material used in many studies, identified as “chlorophyllin”, was quite often, if not always, a copper complex of chlorophyllins (E 141(ii)). In a review, Scotter (2011) stated that: “It is important to consider that despite a joint initiative introduced by the International Union of Pure and Applied Chemistry and the International Union of Biochemistry, a substantial body of long- established trivial names for chlorophyll and its analogues remains in popular use by both the food colour industry and scientific researchers. The term ‘chlorophyllin’ covers a range of compounds identical to, or structurally related to the porphyrins”. There are no data regarding the absorption, distribution, metabolism and excretion (ADME) and toxicity of chlorophyllins (E 140(ii)). From the available data on chlorophylls and their related derivatives, the Panel noted that, in dogs and in humans, the major metabolites of chlorophylls are phaeophytins a and b, with only traces of EFSA Journal 2015;13(5):4085 2 2 Re-evaluation of chlorophyllins (E 140(ii)) as food additives dephytylated metabolites being observed in faeces. Based on these data, the Panel considered that the cleavage of the phytol chain during digestion is unlikely in humans and consequently that chlorophyllins are not metabolites of chlorophylls in humans. Enzymatic hydrolysis of the phytol chain from chlorophylls to yield chlorophyllins can occur only through a chlorophyllase, which is present only in plants and ruminants. Therefore, the Panel concluded that the available data do not support that chlorophyllins are regular metabolites of chlorophylls in humans and it was not possible to support read-across for systemic toxicity data between these two compounds. Some usage data provided to the Panel did not distinguish between chlorophylls (E 140(i)) and chlorophyllins (E 140 (ii)) and therefore, there was uncertainty whether some usage data refer to chlorophylls (E 140(i)) or chlorophyllins (E 140(ii)). Therefore, the exposure to chlorophyllins (E 140(ii)) may possibly be overestimated. Using the “maximum level exposure assessment scenario”, mean exposure to chlorophyllins (E 140(ii)) from their use as food additives ranged from 0.5 mg/kg body weight (bw)/day in the elderly to 12.0 mg/kg bw/day in toddlers. The high exposure to chlorophyllins (E 140(ii)) using this scenario ranged from 1.3 mg/kg bw/day in the elderly to 20.4 mg/kg bw/day in toddlers. Using the refined brand-loyal assessment exposure scenario, mean exposure to chlorophyllins (E 140(ii)) from their use as a food additive ranged from 0.3 mg/kg bw/day in the elderly to 7.2 mg/kg bw/day in toddlers. The high exposure to chlorophyllins (E 140(ii)) using this scenario ranged from 1.0 mg/kg bw/day in the elderly to 16.4 mg/kg bw/day in toddlers. Using the refined non-brand-loyal assessment exposure scenario, mean exposure to chlorophyllins (E 140(ii)) from their use as a food additive ranged from 0.1 mg/kg bw/day in adults and the elderly to 3.1 mg/kg bw/day in toddlers.